肝豆灵通过调控LncRNA H19促进胆汁排铜改善Wilson病肝纤维化OA

Objective:To observe the effects of Gandouling(GDL)on hepatic and serum exosomal long non-coding RNA H19(LncRNA H19)as well as cholestasis in hepatic tissues of copper-loaded rats with Wilson disease,and to explore the mecha-nism of action of GDL ameliorating hepatic fibrosis in Wilson disease.Methods:A total of 60 SD male rats were randomly divided into the normal group(Control),the model group(Model),the GDL low-dose group(GDL-L),the GDL medium-dose group(GDL-M),the GDL high-dose group(GDL-H),and the penicillamine group(PCA),with 10 rats in each group.A copper-loaded Wilson disease liver fibrosis model was constructed by feeding copper(Ⅱ)sulfate pentahydrate(CuSO4·5H2O),and the drug was administered simultaneously during the modeling process.Each treatment group of GDL and the PCA group were administered the corresponding doses of the drugs by gavage,while the Control group and the Model group were administered an equal volume of distilled water by gavage for 30 consecutive days.H&E and Masson staining were used to observe the pathological changes in the liver.ELISA was utilized to measure cholestasis indicators,including γ-glutamyl transferase(GGT),alkaline phosphatase(ALP),total bilirubin(TBIL),and total bile acids(TBA),as well as liver function markers alanine aminotransferase(ALT)and aspartate aminotransferase(AST).Atomic absorption spectrophotometer was used to determine the copper content of bile.RT-qPCR was used to detect the gene expressions of LncRNA H19,cytokeratin 7(CK7),an indicator of bile duct response,and cytokeratin 19(CK19);Western blot was used to observe the protein expressions of CK7 and CK19.Results:Compared to the Control group,the histopathologic pattern of liver tissue in the Model group showed unclear cell nucleus boundaries with lysis and necrosis,and obvious collagen deposition around the central vein and bile ducts of the liver tissue;the levels of ALT,AST,GGT,ALP,TBIL,TBA,and biliary copper were elevated significantly(P<0.01).LncRNA H19,CK7,and CK19 gene ex-pression levels increased significantly(P<0.01).CK7 and CK19 protein levels increased significantly(P<0.01).The degree of hepatic tissue fibrosis was significantly improved after the intervention of GDL and PCA.ALT,AST,GGT,ALP,TBL,TBA,and biliary copper levels were decreased(except for ALT、TBIL and TBA in the GDL-L group)(P<0.01),LncRNA H19,CK7,and CK19 gene expression levels were decreased(P<0.01),and CK7 and CK19 protein levels were decreased(except for CK7 in the GDL-L group)(P<0.05).Conclusion:GDL may promote biliary copper excretion and ameliorate hepatic fibrosis in copper-loaded rats with Wilson disease by regulating the expression of LncRNA H19.